Opioid receptor modulator dosage formulations

ABSTRACT

Abuse deterrent solid dosage formulations containing 5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoic acid, and processes for the preparation and administration of these formulations.

FIELD OF THE DISCLOSURE

The present disclosure relates to oral dosage formulations containing5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid, and processes for the preparation and administration of theseformulations.

BACKGROUND OF THE DISCLOSURE

Delivering an active pharmaceutical ingredient(“5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid”) to a patient requires more than just identifying a molecule andits use. An5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid must be formulated for delivery to a patient and this formulation(in addition to the5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid activity) is evaluated by regulatory agencies such as the US Foodand Drug Administration (FDA) and the European Medicines Agency (EMA).The FDA evaluates the formulation for, among other properties, deliveryproperties, stability, consistency, and manufacturing controls. Animportant factor in determining these properties of a particularformulation is the composition and form of the dosage formulation of the5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid. The formulations for every5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid are different and different formulations containing the same5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid may have very different stability and drug delivery (e.g.,pharmacokinetic) properties.

5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid is an opioid receptor modulator that effects simultaneous agonismof the μ opioid receptor (MOR) and antagonism of the δ opioid receptor(DOR) and may be useful in the treatment and prevention of variousmammalian disease states, for example pain and gastrointestinaldisorders such as diarrheic syndromes, motility disorders includingpost-operative ileus and constipation, and visceral pain includingpost-operative pain, irritable bowel syndrome and inflammatory boweldisorders (for example, see U.S. Pat. No. 7,741,356 to Breslin, et al.,which is incorporated herein in its entirety). Irritable bowel syndromeis a common functional gastrointestinal disorder that affectsapproximately 10-15% of the population in western countries (Lovell etal., Clin Gastroenterol Hepatol 2012;10(7):712-21). Irritable bowelsyndrome is characterized by recurrent abdominal discomfort and painassociated with altered bowel habits (Drossman D A, Gastroenterol2006;130(5):1377-1390). Currently irritable bowel syndrome subtypesinclude diarrhea (IBS-D), constipation (IBS-C), or mixed constipationand diarrhea (IBS-M). Irritable bowel syndrome can negatively impactindividual's quality of life and results in significant direct andindirect costs (Drossman D A. Rome III The Functional GI Disorders.3^(rd) Edition. Lawrence: Allen Press, Inc, 2006). Current safe andeffective pharmacologic treatments for IBS-D are limited and includeantispasmodics, antidepressants, antidiarrheal agents, and alosetron(Brandt et al., Am J Gastroenterol 2009;104(Suppl 1):S1-35).

Opioid receptors, including mu, delta, and kappa are expressed along thegastrointestinal tract and play a key role in regulatinggastrointestinal motility, secretion and visceral sensation (Bagnol etal., Neuroscience 1997;81(2):579-591; Dokray G J, Physiology of EntericNeuropeptides. In: Johnson L R ed. Physiology of the GastrointestinalTract. 3^(rd) ed. New York: Raven, 1994;169-209; Bitar et al., Nature1982;297(5861):72-74). Exogenous opioids reduce gastrointestinal transitthrough activation of MOR and can treat diarrhea in acute situations(Holzer P., Regulatory Peptides 2009;155:11-17). Agents thatsimultaneously activate MOR while antagonizing DOR have differentialgastrointestinal effects and may possess increased analgesic potencycompared to pure MOR agonists (Ananthan S. Opioid ligands with mixed μ/δopioid receptor interactions: an emerging approach to novel analgesics.AAPS Journal 2006;8(1):E118-E125; Dietis et al., British Journal ofAnaesthesia 2009;103(1):38-49). Such a mixed MOR agonist/DOR antagonistprofile may offer an advantage in treating both the diarrhea andabdominal pain associated with IBS-D.

5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid may be particularly useful for reducing pain and diarrhea inpatients with irritable bowel syndrome with diarrhea (IBS-D) withoutconstipating side effects. In vitro, it reduces contractility inintestinal tissue and inhibits neurogenically-mediated secretion (Wadeet al., Br J Pharmacol 2012;167(5):1111-1125). In vivo, it reducesgastrointestinal transit and fecal output in stressed and non-stressedmice over a wide dose-range without fully inhibiting gastrointestinaltransit (ibid.). In contrast, loperamide had a narrow dose range in thesame stressed and non-stressed models and completely prevented fecaloutput in a dose-dependent manner (ibid.).

5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid and methods of making this molecule are disclosed in U.S. Pat. No.7,741,356. Example 9 of U.S. Pat. No. 7,741,356 makes the hydrochloridesalt of5-(1[2-amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-aminol-methyl)-2-methoxy-benzoicacid. Applicants have also discovered a process of making the zwitterionof5-({[2-amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid and two novel crystals of this zwitterions (for example, see U.S.Patent Publication No. 2011/0263868 to Anzalone, et. al., which isincorporated herein in its entirety).

Oral administration of5-({[2-amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid is efficacious in nounalizing gastrointestinal (GI) motility instressed subjects and providing anti-visceral hyperalgesic effects inrats by acting at peripheral opioid receptors in the gastrointestinaltract. It has also been noted that parenteral administration of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid results in CNS-related effects in animal models, which is believedto be due to the mu-opioid receptor (“MOR”) agonist properties.

The recent draft guidance issued for industry by the US Food and DrugAdministration (Food and Drug Administration 2010) for the assessment ofabuse potential of drugs provides general instructions for in vitrolaboratory assessment procedures. This draft guidance states that“[i]nformation should be obtained on how much drug substance might bereleased and any changes that could take place in the rate of release ofthe drug from the drug product if it is misused either intentionally orunintentionally.” The guidance further states that the “effects of pH,temperature, and solvent polarity on disruption of the drug productmatrix should be evaluated. Additional experimental variables mayinclude exposure times to the solvent, agitation, varying the surfacearea (such as from intact to being ground, crushed, or cut into pieces),and ease of crushing tablets or destroying the dosage from matrix.”These guidelines pertain to compounds perceived to have any potentialfor abuse, misuse and/or diversion, which include, but are not limitedto, opioid receptor agonists. The goal is to determine the effects thatcertain formulations may have in limiting or preventing abuse of theactive ingredient, in this instance,5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid, in order to decrease abuse or diversion of the marketed dosageformulations and prevent harm and addiction in the public to the extentpossible.

Therefore an abuse liability assessment of oral dosage formulations of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid was undertaken to identify oral formulation compositions andcharacteristics that may provide effective treatment of opioid receptordisorders while minimizing or elimination potential for abuse ordiversion of these oral formulations.

SUMMARY OF DISCLOSURE

The present inventors have discovered solid oral pharmaceuticalformulations of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid with improved stability and shelf life and unique physico-chemicalfeatures that may deter or limit abuse of the active ingredient ordiversion of the oral formulations. Thus, embodiments provided by thisdisclosure include an abuse deterrent pharmaceutical formulation of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenye-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid.

One embodiment of the disclosure provides a solid pharmaceuticalformulation comprising5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid and an inert ingredient selected from silicified microcrystallinecellulose, colloidal silicon dioxide, crospovidone(polyvinylpolypyrrolidone; highly cross-linked polyvinylpyrrolidone(PVP)), mannitol, and magnesium stearate. In a specific embodiment, thispharmaceutical formulation may be substantially or completely free of aseparate opioid antagonist, such as naloxone. A related embodimentprovides a pharmaceutical formulation consisting of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid and an inert ingredient selected from silicified microcrystallinecellulose, colloidal silicon dioxide, crospovidone, mannitol, andmagnesium stearate.

One embodiment of the disclosure provides a solid pharmaceuticalformulation comprising5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid and inert ingredients including silicified microcrystallinecellulose, colloidal silicon dioxide, crospovidone(polyvinylpolypyrrolidone; highly cross-linked polyvinylpyrrolidone(PVP)), mannitol, and magnesium stearate. In a specific embodiment, thispharmaceutical formulation may be substantially or completely free of aseparate opioid antagonist, such as naloxone. A related embodimentprovides a pharmaceutical formulation consisting of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid, and the inert ingredients silicified microcrystalline cellulose,colloidal silicon dioxide, crospovidone, mannitol, and magnesiumstearate.

One embodiment of the disclosure provides a solid oral dosageformulation comprising5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid and an inert ingredient selected from silicified microcrystallinecellulose, colloidal silicon dioxide, crospovidone(polyvinylpolypyrrolidone; highly cross-linked polyvinylpyrrolidone(PVP)), mannitol, and magnesium stearate. In a specific embodiment, thissolid oral dosage formulation may be substantially or completely free ofa separate opioid antagonist, such as naloxone. A related embodimentprovides a solid oral dosage formulation consisting of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid and an inert ingredient selected from silicified microcrystallinecellulose, colloidal silicon dioxide, crospovidone, mannitol, andmagnesium stearate. In these embodiments, the oral dosage formulationsmay be coated, including sugar coated, gelatin coated, film coated orenteric coated, by standard techniques.

Another embodiment of the disclosure provides an oral tablet formulationcomprising5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid and an inert ingredient selected from silicified microcrystallinecellulose, colloidal silicon dioxide, crospovidone(polyvinylpolypyrrolidone; highly cross-linked polyvinylpyrrolidone(PVP)), mannitol, and magnesium stearate. In a specific embodiment, thisoral tablet formulation may be substantially or completely free of aseparate opioid antagonist, such as naloxone. A related embodimentprovides an oral tablet formulation consisting of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid and an inert ingredient selected from silicified microcrystallinecellulose, colloidal silicon dioxide, crospovidone, mannitol, andmagnesium stearate. In these embodiments, the oral tablet formulationsmay be coated, including sugar coated, gelatin coated, film coated orenteric coated, by standard techniques.

Another embodiment of the disclosure provides a film-coated, oral tabletformulation comprising5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid and an inert ingredient selected from silicified microcrystallinecellulose, colloidal silicon dioxide, crospovidone(polyvinylpolypyrrolidone; highly cross-linked polyvinylpyrrolidone(PVP)), mannitol, and magnesium stearate, and a film coating. In aspecific embodiment, this film-coated, oral tablet formulation may besubstantially or completely free of a separate opioid antagonist, suchas naloxone. A related embodiment provides a film-coated, oral tabletformulation consisting of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid and an inert ingredient selected from silicified microcrystallinecellulose, colloidal silicon dioxide, crospovidone, mannitol, andmagnesium stearate, and a film coating. In these embodiments, the filmcoating may be an aqueous film coating.

A specific embodiment is an abuse deterrent, mono-phasic pharmaceuticalcomposition suitable for single dose administration for treating orameliorating a condition mediated by an opioid receptor consistingessentially of about 20 mg/dose to about 200 mg/dose of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid, from about 60-80% by weight of silicified microcrystallinecellulose, from about 2-8% by weight of colloidal silica, from about50-90% by weight of mannitol, from about 20-50% by weight ofcrospovidone, and from about 2-8% by weight of magnesium stearate.Another embodiment provided by this disclosure is a method of treatingor ameliorating a condition mediated by an opioid receptor byadministering5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenye-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid in a solid oral formulation of this disclosure to a subject in needof such treatment. In a specific embodiment, this administration may bemade in the absence of the separate or concurrent administration of anopioid antagonist, such as naloxone. In specific embodiments, thesemethods may include the administration of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid to the subject in an amount between 20 mg and 200 mg.

In specific embodiments, these methods may include the administration of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid to the subject in an amount between about 10 mg and about 125 mg.In specific embodiments, these methods may include the administration of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid to the subject in an amount between about 50 mg and about 100 mg.In specific embodiments, these methods may include the administration of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenye-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid to the subject in an amount of about 75 mg. In specificembodiments, these methods may include the administration of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid to the subject in an amount of about 100 mg.

In specific embodiments, these methods may include the administration of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid in a formulation of this disclosure to the subject between twoadministrations per day and eight administrations per day. In specificembodiments, these methods may include the administration of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid in a formulation of this disclosure to the subject between twoadministrations per day and six administrations per day. In specificembodiments, these methods may include the administration of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid in a formulation of this disclosure to the subject between twoadministrations per day and four administrations per day. In specificembodiments, these methods may include the administration of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid in a formulation of this disclosure to the subject on a twice-dailydosing regimen. In specific embodiments, these methods may include theadministration of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-ye-ethyl]-amino}-methyl)-2-methoxy-benzoicacid in a formulation of this disclosure to the subject on a once-dailydosing regimen.

This Summary of the Disclosure is neither intended nor should it beconstrued as being representative of the full extent and scope of thepresent disclosure. Moreover, references made herein to “the presentdisclosure,” or aspects thereof, should be understood to mean certainembodiments of the present disclosure and should not necessarily beconstrued as limiting all embodiments to a particular description. Thepresent disclosure is set forth in various levels of detail in theSummary of the Disclosure as well as in the attached drawings and theDescription of Embodiments and no limitation as to the scope of thepresent disclosure is intended by either the inclusion or non-inclusionof elements, components, etc. in this Summary of the Disclosure.Additional aspects of the present disclosure will become more readilyapparent from the Description of Embodiments, particularly when takentogether with the drawings.

BRIEF DESCRIPTION OF DRAWINGS

FIG. 1 depicts the manufacturing process for 75-mg and 100-mg oraltablets of the present disclosure.

FIG. 2, shows representative time course plots of cumulative percentrecovery of active ingredient under different extraction conditions: A,ground vs. whole tablet; B, 25° C. vs. 95° C.; C, water vs. 0.1M HCl.

DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

The present disclosure is drawn to solid dosage formulations containing5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid that deter or minimize the abuse or diversion of theseformulations, as well as processes for the preparation andadministration of these formulations.

For the purposes of this disclosure, reference to“5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid” also means“5-[[[-2-amino-3-[4-(aminocarbonyl)-2,6-dimethylphenyl]-1-oxopropyl][-1-(4-phenyl-1H-imidazol-2-yl)ethyl]amino]methyl]-2-methoxybenzoicacid”, and it is intended that the two chemical names can be usedinterchangeably. Reference to the “active ingredient” includes5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid, and pharmaceutically acceptable enantiomers, diastereomers,racemates, zwitterions, and salts thereof.

Because5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]amino}-methyl)-2-methoxy-benzoicacid includes at least two chiral centers, it may exist asdiastereomers. These isomers may be separated by conventional techniquessuch as preparative chromatography and may be prepared in racemic formor as individual diasteromers by either stereospecific synthesis or byresolution. The compounds may, for example, be resolved into theircomponent diasteromers by standard techniques, such as the formation ofstereoisomeric pairs by salt formation with an optically active acid,such as (−)-di-p-toluoyl-D-tartaric acid and/or(+)-di-p-toluoyl-L-tartaric acid followed by fractional crystallizationand regeneration of the free base. The compounds may also be resolved byformation of stereoisomeric esters or amides, followed bychromatographic separation and removal of the chiral auxiliary.Alternatively, the compounds may be resolved using a chiral HPLC column.It is to be understood that all stereoisomers, racemic mixtures,diastereomers and enantiomers are encompassed within the scope of anyreference to“5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid” or the “active ingredient” in this disclosure.

For example, reference to“5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid” or the “active ingredient” specifically includes:

-   -   a)        5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenye-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoic        acid,    -   b)        5-({[(2S)-2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoic        acid,    -   c)        5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[(1S)-1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoic        acid, and/or    -   d)        5-({[(2S)-2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[(1S)-1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoic        acid, as well as    -   e) a hydrobromic, hydriodic, perchloric, sulfuric, nitric,        phosphoric, acetic, propionic, glycolic, lactic, succinic,        maleic, fumaric, malic, tartaric, citric, benzoic, mandelic,        methanesulfonic, hydroxyethanesulfonic, benzenesulfonic, oxalic,        pamoic, 2-naphthalenesulfonic, p-toluenesulfonic,        cyclohexanesulfamic, salicylic, saccharinic or trifluoroacetic        acid salt of any one of these compounds, or    -   f) a benzathine, chloroprocaine, choline, diethanolamine,        ethylenediamine, meglumine, procaine, aluminum, calcium,        lithium, magnesium, potassium, sodium, or zinc salt of any one        of these compounds.

Similarly, reference to“5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenye-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid” or the “active ingredient” specifically includes a compound havingthe chemical structure:

as well as a hydrobromic, hydriodic, perchloric, sulfuric, nitric,phosphoric, acetic, propionic, glycolic, lactic, succinic, maleic,fumaric, malic, tartaric, citric, benzoic, mandelic, methanesulfonic,hydroxyethanesulfonic, benzenesulfonic, oxalic, pamoic,2-naphthalenesulfonic, p-toluenesulfonic, cyclohexanesulfamic,salicylic, saccharinic or trifluoroacetic acid salt, or a benzathine,chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine,procaine, aluminum, calcium, lithium, magnesium, potassium, sodium, orzinc salt of any one of these chemical structures.

As used in this disclosure, the terms “subject,” “patient,”“individual,” etc. are not intended to be limiting and can be generallyinterchanged. That is, an individual described as a “patient” does notnecessarily have a given disease, but may be merely seeking medicaladvice.

As used herein, the terms “treat” and “prevent” are not intended to beabsolute terms. Treatment can refer to any delay in onset, reduction inthe frequency or severity of symptoms, amelioration of symptoms,improvement in patient comfort and/or gastrointestinal function, etc.The effect of treatment can be compared to an individual or pool ofindividuals not receiving a given treatment, or to the same patientprior to, or after cessation of, treatment.

The term “prevent” or “ameliorate” refers to a decrease in theoccurrence of opioid receptor disease or disorder symptoms in a patient.The prevention or amelioration may be complete (no detectable symptoms)or partial, such that fewer symptoms are observed than would likelyoccur absent treatment.

The phrase “opioid receptor related disorders” refers to variousmammalian disease states including, for example, pain andgastrointestinal disorders such as diarrheic syndromes, motilitydisorders including post-operative ileus and constipation, and visceralpain including post-operative pain, irritable bowel syndrome andinflammatory bowel disorders, as described in greater detail in U.S.Pat. No. 7,741,356 to Breslin, et al., which is incorporated herein inits entirety.

The term “abusive” or “abusive manner” refers to uses of theformulations of this disclosure beyond oral administration, such as byinjecting or snorting.

The term “abuse deterrent”, as used herein, refers to a formulation ofthe present invention which possesses physico-chemical characteristicsthat allow the therapeutic use of the opioid receptor modulator activeingredient by oral administration to a subject in need of such treatmentwith very limited potential for abuse or misuse of the formulation,i.e., by extracting and ingesting the active ingredient by snorting orinjection.

The term “therapeutically effective amount,” as used herein, refers tothat amount of the therapeutic agent sufficient to treat or amelioratethe disease or disorder, as described above. For example, for the givenparameter, a therapeutically effective amount will show an increase ordecrease of at least about 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%,70%, 80%, 90%, or 100%. Therapeutic efficacy can also be expressed as“-fold” increase or decrease. For example, a therapeutically effectiveamount can have at least a 1.2-fold, 1.5-fold, 2-fold, 5-fold, or moreeffect over a control value. Therapeutic efficacy related to irritablebowel syndrome, preferably diarrhea-predominant irritable bowelsyndrome, can be expressed in measurements of worst abdominal pain (WAP)or stool consistency score (Bristol Stool Scale or BSS).

The term “solid dosage formulation” as used herein includes tablets,capsules, pills and like and may be present as conventional orextended-release compositions.

The terminology used herein is for describing particular embodiments andis not intended to be limiting. As used herein, the singular forms “a,”“and” and “the” include plural referents unless the content and contextclearly dictate otherwise. Thus, for example, a reference to “a marker”includes a combination of two or more such markers. Unless definedotherwise, all scientific and technical terms are to be understood ashaving the same meaning as commonly used in the art to which theypertain. For the purposes of the present disclosure, the following termsare defined below.

Also, the use of “or” means “and/or” unless stated otherwise. Similarly,“comprise,” “comprises,” “comprising,” “include,” “includes,” and“including” are interchangeable and not intended to be limiting.

It is to be further understood that where descriptions of variousembodiments use the term “comprising,” those skilled in the art wouldunderstand that in some specific instances, an embodiment can bealternatively described using language “consisting essentially of” or“consisting of.”

In some embodiments related to the treatment of opioid receptor relateddisorders, the dose of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid that can be incorporated into the formulations of the presentdisclosure depends on the desired treatment dosage to be administeredand can range from about 20 mg to about 200 mg of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid. In other embodiments, the dose of the active ingredient can rangefrom about 10 mg to about 125 mg. In some embodiments, the dose of theactive ingredient in the formulations of this disclosure is betweenabout 10 mg and about 200 mg, e.g., 10 mg, 15 mg, 20 mg, 25 mg, 30 mg,35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85mg, 90 mg, 95 mg, 100 mg, 105 mg, 110 mg, 115 mg, 120 mg, 125 mg, 130mg, 135 mg, 140 mg, 145 mg, 150 mg, 155 mg, 160 mg, 165 mg, 170 mg, 175mg, 180 mg, 185 mg, 190 mg, 195 mg, or 200 mg of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenye-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid. In one embodiment, the dose is about 75 mg of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid. In another embodiment, the dose is about 100 mg of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid. In another embodiment, the dose is about 50 mg of 5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid. In yet another embodiment, the dose is about 150 mg of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino} -methyl)-2-methoxy-benzoic acid. In yet another embodiment, the doseis about 37.5 mg of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid.

Dose proportionality occurs when increases in the administered dose areaccompanied by proportional increases in a pharmacokinetic parameter.The dosage formulations of the present disclosure are preferablydesigned as a dose proportional formulation in the range of 25 mg to 100mg of the active ingredient5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid.

Abuse deterrent formulations of the present invention include solidpharmaceutical dosage formulations containing:

-   -   from about 5-20% by weight of        5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoic        acid, preferably from about 10-15% by weight of        5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoic        acid, preferably about 12.5% by weight of        5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoic        acid;    -   from about 60-80% by weight of silicified microcrystalline        cellulose (USP Silicified Microcrystalline Cellulose; intimately        associated microcrystalline cellulose and colloidal silicon        dioxide particles; in a preferred example, high density        silicified microcrystalline cellulose “HD-90”); preferably from        about 65-75% by weight of silicified microcrystalline cellulose,        preferably about 71% by weight of silica fine microcrystalline        cellulose;    -   from about 0.45-1.0% by weight of a glidant, e.g., colloidal        silica, preferably from about 0.55-0.95% by weight of colloidal        silica, in one preferred embodiment, about 0.65%-0.85% by weight        of colloidal silica, in another preferred embodiment, about        0.75% by weight of colloidal silica;    -   from about 1-20% by weight of mannitol (mannitol, USP),        preferably from about 5-15% by weight of mannitol, in one        preferred embodiment, about 7.5%-12.5% by weight of mannitol, in        another preferred embodiment, about 10% by weight of mannitol;    -   from about 2-8% by weight of crospovidone (highly cross-linked        modification of polyvinylpyrrolidone (PVP)), preferably from        about 3-7% by weight of crospovidone, in one preferred        embodiment, about 4-6% by weight of crospovidone, in another        preferred embodiment, about 5% by weight of crospovidone; and,    -   from about 0.45-1% by weight of magnesium stearate (Magnesium        Stearate USP), preferably from about 0.55-0.95% by weight of        magnesium stearate, in one preferred embodiment, about        0.65%-0.85% by weight of magnesium stearate, in another        preferred embodiment, about 0.75% by weight of magnesium        stearate.

In some embodiments, the abuse deterrent solid pharmaceutical dosageformulations are coated with a film coating. In some embodiments, thecoating is a water-soluble, pH-independent film coating. Such coatingallows for immediate disintegration for fast, active release of thecontents of the solid dosage formulation. One such commerciallyavailable coating useful in the solid dosage formulations of the presentdisclosure is Opadry®, and preferably Opadry II®. In specificembodiments, the abuse deterrent solid pharmaceutical dosageformulations include a film coating present in an amount of from about0.5-5.5% by weight film coating, preferably from about 1-5% by weightfilm coating, in one preferred embodiment, about 2-4% by weight filmcoating, in another preferred embodiment, about 3% by weight filmcoating. In some instances, opioid receptor agonist drugs have beenformulated and marketed with an opioid antagonist (such as the opioidantagonist naloxone), to render the formulation abuse resistant. Thepresent invention is also based, at least in part, on the surprisingdiscovery, that the formulations of the present disclosure have abusedeterrent characteristics. For example, such formulation isnon-tamperable thereby limiting ease of isolation and purification of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid from the formulation. Thus, in specific embodiments of the presentdisclosure, the formulations are substantially free of naloxone. Inrelated embodiments of the present disclosure, the formulations arecompletely free of naloxone. Such formulations are advantageous becausethey can provide effective abuse deterrence in the absence of naloxone.

The formulations can also optionally include an inert pharmaceuticallyacceptable dissolution-rate-modifying agent, a pharmaceuticallyacceptable plasticizer, a pharmaceutically acceptable coloring agent(e.g., FD&C Blue #1), a pharmaceutically acceptable opacifier (e.g.,titanium dioxide), pharmaceutically acceptable anti-oxidant (e.g.,tocopherol acetate), a pharmaceutically acceptable preservative,flavorants (e.g., saccharin and peppennint), neutralizing agents (e.g.,sodium hydroxide), buffering agents (e.g., monobasic, or tribasic sodiumphosphate), or combinations thereof. Preferably, these components areindividually present at no more than about 1% of the final weight of theformulation, but the amount may vary depending on the other componentsof the formulation.

For preparing formulations of the present disclosure, such as tablets,5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid is mixed with one or more pharmaceutical excipients to form a solidpreformulation composition containing, in preferred embodiments, ahomogeneous mixture of the excipient(s) with the active ingredient. Whenreferring to these preformulation compositions as “homogeneous,” it ismeant that the active ingredient are dispersed evenly throughout thecomposition so that the composition may be readily subdivided intoequally effective unit dosage forms such as tablets or capsules. Thissolid preformulation is then subdivided into unit dosage forms of thetype described above containing from, for example, about 10 to about 200milligrams of the active ingredient.

According to one embodiment, a solid dosage formulation of the presentdisclosure is prepared by:

-   -   i) blending        5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoic        acid with the pharmaceutically acceptable inert excipients:        silicified microcrystalline cellulose, mannitol, colloidal        silica and crospovidone;    -   ii) comilling the above blend with the addition of magnesium        stearate;    -   iii) compressing the dry blend into suitably sized tablets, or        filling into capsules.

According to another embodiment, a solid dosage formulation is preparedusing direct compression, by:

-   -   i) blending        5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoic        acid with the pharmaceutically acceptable inert excipients        silicified microcrystalline cellulose, mannitol, colloidal        silica, and crospovidone;    -   ii) lubricating the blend by the addition of magnesium stearate;    -   iii) compressing the blend into suitably sized tablets.

According to another embodiment, capsules may be formulated by:

-   -   i) blending        5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-ye-ethyl]-amino}-methyl)-2-methoxy-benzoic        acid with the pharmaceutically acceptable inert excipients        silicified microcrystalline cellulose, mannitol, colloidal        silica, and crospovidone;    -   ii) lubricating the blend by the addition of magnesium stearate,    -   iii) filling the blend into capsule shells.

According to another embodiment, capsules may also be formulated by:

-   -   i) blending        5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenye-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoic        acid with the pharmaceutically acceptable inert excipients        silicified microcrystalline cellulose, mannitol, colloidal        silica, and crospovidone;    -   ii) comilling the above blend,    -   iii) adding magnesium stearate to lubricate the blend,    -   iv) filling the blend into capsule shells.

According to another embodiment, capsules may also be formulated by:

-   -   i) blending        5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoic        acid with the pharmaceutically acceptable inert excipients        silicified microcrystalline cellulose, mannitol, colloidal        silica, and crospovidone;    -   ii) comilling the above blend,    -   iii) adding magnesium stearate to lubricate the blend,    -   iv) compressing the blend into tablets.

In these embodiments of preparing the dosage formulations of thisdisclosure, the tablets or capsules may be film coated and/or packagedin bulk or unit-dosage packaging (i.e. blister packaging).

The active ingredient in the formulations of the present disclosure areuseful opioid receptor modulators. In particular, certain compounds areopioid receptor agonists useful in the treatment or amelioration ofconditions such as pain and gastrointestinal disorders. Examples of painintended to be within the scope of the present invention include, butare not limited to, centrally mediated pain, peripherally mediated pain,structural or soft tissue injury related pain, pain related toinflammation, progressive disease related pain, neuropathic pain andacute pain such as caused by acute injury, trauma or surgery, andchronic pain such as caused by neuropathic pain conditions, diabeticperipheral neuropathy, post-herpetic neuralgia, trigeminal neuralgia,post-stroke pain syndromes or cluster or migraine headaches. The activeingredient in the formulations of the present disclosure are preferablyuseful for treating or ameliorating abdominal pain. Examples ofgastrointestinal disorders intended to be within the scope of thisinvention include, but are not limited to, diarrheic syndromes, motilitydisorders such as irritable bowel syndrome includingdiarrhea-predominant, constipation-predominant or alternating irritablebowel syndrome, and visceral pain and diarrhea associated withinflammatory bowel disease including ulcerative colitis and Crohn'sdisease.

Examples of gastrointestinal disorders where-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid may be useful include constipation-predominant irritable bowelsyndrome, post-operative ileus and constipation, including but notlimited to the constipation associated with treatment of chronic painwith opiates. Modulation of more than one opioid receptor subtype isalso useful as follows: a compound that is a mixed mu OR agonist anddelta OR antagonist could have antidiarrheal properties without beingprofoundly constipating. A compound that is a mixed mu OR agonist anddelta OR agonist are useful in cases of severe diarrhea that arerefractory to treatment with pure mu OR agonists, or has additionalutility in treating visceral pain associated with inflammation anddiarrhea.

The daily dose of a solid dosage formulation of the present disclosuremay be a dose that is therapeutically effective for the treatment ofirritable bowel syndrome, preferably diarrhea-predominant irritablebowel syndrome, wherein a patient has an average decrease of daily WAPscores from the patient's baseline WAP (for example a baseline WAP scoreof ≧3.0 (on a 0-10 numerical rating scale, where 0 indicates no pain and10 worst pain imaginable) of about ≧10%, preferably a decrease in WAPscore of about ≧20%, more preferably a decrease in WAP score of about≧30%. The daily dose of a dosage formulation of the present disclosuremay be a dose that is therapeutically effective for the treatment ofirritable bowel syndrome, preferably diarrhea-predominant irritablebowel syndrome, wherein a patient achieves an average BSS score (wherein1 equals hard, lumpy stools and 7 equals watery, liquid stools) ofbetween 2 and 5, preferably 3 or 4.

The daily dose of a solid dosage formulation of the present disclosuremay be varied over a wide range from about 20 mg to about 7000 mg of theactive ingredient per adult human per day; preferably the dose will bein the range of from about 50 mg to about 2100 mg of the activeingredient per adult human per day. For oral administration, theformulations are preferably provided in the form of film-coated tabletscontaining about 75, about 100 or about 200 milligrams, more preferablyabout 75 or about 100 milligrams, of the active ingredient for thesymptomatic adjustment of the dosage to the subject to be treated.Advantageously, dosage formulations of the present disclosure may beadministered in a single daily dose or the total daily dosage may beadministered in divided doses of two, three or four times daily. Ifadministered twice daily, each dose may be administered at about 6 hoursto about 12 hours apart per day, preferably about 8 hours to about 12hours apart per day, preferably about 10 hours to about 12 hours apartper day, preferably about 10 hours apart per day, or preferably about 12hours apart per day. In these preferred embodiments, the total dailydose is administered twice daily.

In certain embodiments, these methods may include the administration of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid in a formulation of this disclosure to the subject on anevery-other-day (qod) dosing regimen, or on a thrice-weekly dosingregimen, or on a twice-weekly dosing regimen, or on a once-weekly dosingregimen.

In certain embodiments, these methods may include the administration of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]4[-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid in a formulation of this disclosure to the subject on atwice-monthly dosing regimen, or on a once-monthly dosing regimen.

In certain embodiments, these methods may include the administration of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid in a formulation of this disclosure to the subject on an as-needed(prn) dosing regimen.

Thus, an embodiment provided by this disclosure is a method of treatingor ameliorating an opioid receptor related disorder by administering aformulation of this disclosure to a subject in need of such treatment.In one embodiment, this administration may be made in the absence of theseparate or concurrent administration of the opioid antagonist naloxone.In specific embodiments, these methods may include the administration of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid to the subject in an amount between about 20 mg and about 200 mg,preferably about 75, about 100 or about 200 milligrams, more preferablyabout 75 or about 100 milligrams.

In another embodiment, this administration may be made in the absenceof, prior to or concurrent with, the administration of food. In oneembodiment,5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid is administered to the subject with one or more daily meals, morepreferably with breakfast and dinner.

Another embodiment of the disclosure relates to any of the formulationsof this disclosure for use in the treatment or amelioration of acondition mediated by an opioid receptor, for example, any pain andgastrointestinal disorders disclosed herein, preferably such asdiarrheic syndromes, motility disorders including post-operative ileusand constipation, and visceral pain including post-operative pain,irritable bowel syndrome and inflammatory bowel disorders.

Another embodiment of the disclosure relates to the use of any of theformulations of this disclosure in the preparation of a medicament forthe treatment or amelioration of a condition mediated by an opioidreceptor, for example pain and gastrointestinal disorders disclosedherein, such as diarrheic syndromes, motility disorders includingpost-operative ileus and constipation, and visceral pain includingpost-operative pain, irritable bowel syndrome and inflammatory boweldisorders.

The disclosure now being generally described will be more readilyunderstood by reference to the following examples, which are includedmerely for the purposes of illustration of certain aspects of theembodiments of the present disclosure. The examples are not intended tolimit the disclosure, as one of skill in the art would recognize fromthe above teachings and the following examples that other techniques andmethods can satisfy the claims and can be employed without departingfrom the scope of the claimed disclosure.

EXAMPLES Example 1 Description and Composition of Two Formulations ofDisclosure Description of the Dosage Form

The active ingredient,5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid was formulated as 75-mg and 100-mg film-coated tablets. Theformulation was composed of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid and the inactive components listed in Table 1.

Composition

TABLE 1 Composition of Tablets, 75-mg and 100-mg Strength (label claim)75 mg 100 mg Component and Quality Standard (and Quantity QuantityGrade) Function per mg % per mg % Active ingredient Active 75 12.5 10012.5 Silicified Microcrystalline cellulose (HD90) Filler 426 71.0 56871.0 (NF, Ph. Eur.) Colloidal silica (NF, Ph. Eur.) Glidant 4.5 0.75 60.75 Mannitol (USP, Ph. Eur.) Filler 60 10.0 80 10.0 Crospovidone (PolyPXL10) (NF, Ph. Eur.) Disintegrant 30 5.0 40 5.0 Magnesium stearate (NF,Ph. Eur.) Lubricant 4.5 0.75 6 0.75 Nominal Tablet Weight — 600 100 800100 Opadry II 85F18422 Film coat 18 3.0 24 3.0 (Company Specification)Purified water, USP Film coat —^(a) — —^(a) — solvent Total — 618 — 824— ^(a)Removed during processing

Example 2 Description of one Manufacturing Process (and ProcessControls)

The flow chart for the method of manufacture for the 75- and 100-mg oraltablets is presented in FIG. 1.

Description of the Manufacturing Process of FIG. 1.

-   -   1. Screen silicified microcrystalline cellulose, mannitol,        crospovidone and colloidal silica through a 20 mesh screen and        magnesium stearate through a 30 mesh screen.    -   2. Transfer the following into a 650-L tote bin: half of the        silicified microcrystalline cellulose, all of the active        ingredient, mannitol, colloidal silica, crospovidone, and the        remaining half of silicified microcrystalline cellulose. Blend        at 12 rpm for 10 minutes. Add the magnesium stearate, blend at        12 rpm for 5 minutes and sample.    -   3. Compress the tablets using a Stoke 34D tablet press or        similar with a speed of 35-45 rpm. Collect samples throughout        the compression run.    -   4. Prepare the coating suspension by dispersing the Opadry II        85F18422 in water and mixing. Apply the coating suspension with        a spray gun at 300 g/min in a 48 inch Accela Cota coater with        following parameters:

TABLE 2 Tablet Coating Parameters Coating Parameter Setting No. ofBaffles 4 Number of Spray Guns 4 Spray Apparatus Schlick (Module9347-1535 with pattern and atomization adjustments) Nozzle Diameter/AirCap 1.2 mm/Air Cap 4 mm Delivery System Peristalic Pump Gun to BedDistance^(a) 9″ (8-11″) Set Point Delivery Rate^(a) 300 ± 50 g/min/4Guns Pan Speed^(a) 4.5 (4-8) rpm Atomizing Air^(a) 30 ± 5 psi PatternAir^(a) 25 ± 5 psi Inlet Air Temperature^(a) 60 ± 10° C. Air Volume(Inlet)^(a) 1500 ± 500 cfm Guideline only - adjust as required toachieve a suitable coating process

-   -   5. Cool the tablets and store in tared, labeled high density        polyethylene (HDPE) containers lined with double polyethylene        bags with twist ties.    -   6. Prepare final packaging in ACLAR blisters or HDPE bottles.

Example 3 Abuse Liability Assessment of Oral Formulation

An abuse liability assessment was undertaken in rhesus monkeys todetermine the doses and systemic exposure levels following acuteintravenous administration of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid that would provide a discriminative stimulus inmorphine-conditioned monkeys and positive reinforcing effects(self-administration) in heroin-conditioned monkeys. The drugdiscrimination studies revealed that morphine-trained monkeysdiscriminated between saline at an IV dose of ≧10 mg/kg of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid. Self-administration studies revealed that in monkeys conditionedto self-administer heroin,5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenye-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid (3.2 mg/kg/IV infusion) provided reinforcement forself-administration. However, 1 mg/kg did not produce a signal in eithermorphine- or heroin-conditioned primates.

Thus, at least in animals, oral administration of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid does not produce CNS effects that are prototypic of abused drugs.However, parenteral administration does appear to produce these effects.Typically, when this pattern of effects is observed in animals, it isrecommended to conduct a human laboratory abuse potential study usingboth the therapeutic route (i.e., oral), as well as the IV route, whichwould represent a “worst case scenario.”

In addition the recommendation to conduct in vitro studies to determinethe ease and feasibility of preparing5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid for abuse through alternative routes of administration such asinjection or snorting (e.g., in vitro extractability/tamperabilitystudies). This is consistent with the draft “Guidance for Industry:Assessment of Abuse Potential of Drugs” (FDA, 2010). This examplediscloses the findings of such in vitro extractability/tamperabilitystudies.

Based on the FDA guidance, a series of in vitro laboratory assessmentstudies were designed to explore the abuse potential of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenye-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid. Specific areas that were identified for laboratory assessment wereas follows:

-   -   Physical manipulations and pretreatment effects    -   Aqueous and organic solvent extractions    -   Syringeability assessments    -   Simulated smoking assessments

Laboratory experiments were targeted toward outcomes that could producetampered product suitable for administration by alternate routes ofadministration including injection, intranasal administration andsmoking. Experiment design began with a consideration of the physicaland chemical properties of the5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid and formulation excipients. The solubility profile of the5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid has been characterized as ranging from “slightly soluble” (1-10mg/mL) in water to “sparingly soluble” 10-33 mg/mL in acidic pH2buffered solutions to “freely soluble” (100-1000 mg/mL) in 0.1N NaOHsolution. Consequently, a core group of laboratory assessments weredirected toward experiments that characterized the “extractability” ofthe5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid from the formulation matrix. The formulation tested was the 75 mgand 100 mg tablets described in Example 1, Table 1.

The first phase of the study consisted of assessing the formulation forease of physical manipulation (e.g., crushing, film coating removal, andpretreatments), extractability with selected aqueous and organicsolvents, effects of filtration on assay results, simulated smokingexperiments, and syringeability.

It was determined that first phase assessments of physical manipulations(cutting/crushing/grinding assessments, pretreatment by freezing andheating, and coating removal), and simulated smoking experimentsprovided a clear and complete body of data on these topics and nofurther assessments were needed. The first phase assessments ofextraction provided a basis for a second, formal phase study ofextractions from this formulation.

All assessments of these tablet formulations were performed with the 100mg5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid dosage tablet (total tablet weight=824 mg) with the exception ofphysical manipulation experiments and the Time Course study which wereperformed with the 75 mg strength tablet. The two dosage strengths aredose proportional in relation to excipients (common blend) and have thesame coating.

To assure production of reliable, accurate data, the experimental designof laboratory protocols included the following elements: use ofsufficient replicates to assess method variability; inclusion ofcontrols for comparison where appropriate; investigation over a broadrange of chemical and physical conditions; verification of analyticalmethods; and use of independent laboratories to whom validatedmethodologies had been transferred.

A. Physical Manipulations and Pretreatments

The ease of cutting and crushing of the tablets was assessed with arange of readily available household items including razor blades,spoons, pliers, tablet crushers, hammer, rolling pin, and mortar andpestle. The effect of tablet pretreatment by freezing at −20° C. orheating at 100° C. was also assessed. Coating removal was assessed byrubbing tablets with wet paper towels.

Result Summary and Discussion

Although the tablets were slightly difficult to “crack”, crushing couldreadily be accomplished with a variety of common household tools.Freezing or oven heating of the tablets did not affect tablet“crushability.” Coating removal could be accomplished easily with a wetpaper towel.

B. Extractions

Solvent Extraction

Experiments designed to simulate preparation of the tablets forinjection were conducted by extracting a single powdered or intacttablet at 25° C. with 10 mL of solvent for 15, 30 and 60 minutes withaqueous- and organic-based solvents (water, 0.1M HCl, ethanol, hexane,pH 2.0 buffer, pH 4.0 buffer, pH 7.0 buffer, pH 10.0 buffer, saline, 10%ethanol, acetone, and isopropyl alcohol). The water and ethanolextractions were repeated at 95° C.

Result Summary and Discussion

Percent recoveries of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid for solvent extractions at 25° C. were variable with solvent type.Aqueous-based solvents were more efficient than non-polar organicsolvents (e.g., hexane). Acidic and basic solvents were more efficientthan water or ethanol. Unusual high recoveries of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid were observed unexpectedly for isopropyl alcohol (80% at 60minutes/25° C., ground tablet) and water (118% at 60 minutes/95° C.). Itwas noted that these experiments were conducted with unfiltered aliquotsleading to the suspicion that un-dissolved particles of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid were being transferred and dissolved in the HPLC media duringanalysis, thereby falsely raising the true percent of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid undergoing dissolution (extraction).

Effects of Filtration on5-({[2-Amino-3-(4-Carbamoyl-2,6-Dimethyl-Phenyl)-Propionyl]-[1-(4-Phenyl-1H-Imidazol-2-Yl)-Ethyl]-Amino}-Methyl)-2-Methoxy-BenzoicAcid Extractability

With the suspicion that unfiltered extractions might be a combination oftrue extracted (dissolved)5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid and suspensions of undissolved material, additional extractionswere performed to determine the effect of filtration. Extracts werefirst filtered through 0.45 μm PTFE filters prior to HPLC analysis.Single whole and ground tablets were extracted at 25° C. with 10 mLsolvent for 15, 30 and 60 minutes with water, acetone, isopropylalcohol, ethanol, pH 2.0 buffer, pH 4.0 buffer, pH 7.0 buffer, pH 10.0buffer.

Result Summary and Discussion

Addition of the filtration step reduced recoveries for all solvents. Thegreatest decreases were observed with the organic solvents. A comparisonof the percentage of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid extracted for ground tablets at 60 minutes (25° C. with 10 mLsolvent) is shown in Table 3, below. Clearly, undissolved particles ledto falsely increased estimates of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid recovery and considerable scatter in individual determinations.

TABLE 3 Comparison of the percentage of 5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoic acid extracted from groundtablets % Recovery, % Recovery, Solvent Unfiltered Filtered Water 63 38Acetone 30 2 Isopropyl Alcohol 80 0 Ethanol 52 5 pH 2 Buffer 103 36 pH 4Buffer 58 12 pH 7 Buffer 81 22 pH 10 Buffer 100 33

C. Syringeability

Experiments were conducted to determine the syringeability of tabletextracts. These assessments were performed on the remainder of solutions(following aliquot removal for determination of percentage of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid recovery) produced in the first phase extractions at 25° C.Solutions were aspirated into disposable syringes equipped with 25 gaugeneedles.

Result Summary and Discussion

All solutions were successfully loaded into disposable syringes equippedwith a 25 gauge needle indicating that extracted solutions offered noresistance to syringeability.

D. Simulated Smoking Assessment

Simulated smoking of the active ingredient was assessed by heatingground or intact tablets in a test tube fitted with an apparatus thatcollected vaporized5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid. As heat from a heating block was applied to the sealed test tube,air was drawn through the tube and over the surface of the heatedproduct. The air exited through a collector cartridge (C18 Sep-Pakcartridge) situated over the heated product. Product was heated to 225°C. until the material was charred (usually approximately 5 minutes).Samples were heated for a total of 10 minutes. In addition, pure5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid was treated in the same manner. The capacity for the apparatus tocollect vaporized5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid was verified with a series of tests involving passing a standardsolution of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid through the cartridge, and testing for “breakthrough” with seriallyconnected cartridges. Recoveries of standard amounts of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid reference material trapped by the collector were essentiallyquantitative. Heating ground and intact tablets with a torch toextremely high temperatures was also attempted followed by collection ofvaporized material with a bubbler collector.

Result Summary and Discussion

Browning and charring was evident when tablets and ground product wereheated to 225° C. in the heating block or to extreme heat with a torch.No detectable5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid was vaporized from ground or intact tablets when heated by eithermethod. Only extremely minor traces of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1-(4-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid were vaporized when pure5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid was heated under the same conditions. Mass balance analysis wasattempted by analyzing the remaining5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid in the heated test tubes. Generally,5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid recoveries from the heated tube were low likely indicating thatheating under these conditions produced thermal decomposition of the5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid. Consequently, it seems safe to conclude that smoking is not aviable route of administration for the5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid.

E. Aqueous Solvent Extraction

Small volume extractions were conducted with various household solventsand buffers to simulate extraction procedures that might be used forpreparation of solutions for injection. Extractions were performed onsix replicates of ground and intact tablets at 25° C. and 95° C. with 10mL of solvent for 10 minutes. The extractions were shaken on an orbitalshaker at 100 rpm, and then aliquots were removed and filtered with 0.45μm PTFE filters for HPLC analysis. The solvents utilized in theseassessments were water, saline, vinegar, 0.1M HCl, 10% ethanol, 40%ethanol, pH 2.0 buffer, pH 4.0 buffer, pH 7.0 buffer, and pH 10.0buffer. Results were expressed in Table 4 below as % recovery (% labelclaim) and as concentration (mg/mL).

Result Summary and Discussion

TABLE 4 Summary of mean results (n = 6) is shown in the table below.Conc. Conc. ° C. % Recovery, % Recovery, (mg/mL), (mg/mL), Solvent TempGround Intact Ground Intact Water 25 22.1 16.4 2.2 1.6 95 39.9 38.7 4.03.9 Saline 25 13.7 17.0 1.4 1.7 95 38.9 61.4 3.9 6.1 Vinegar 25 80.024.2 8.0 2.4 95 77.7 77.9 7.8 7.8 0.1M HCl 25 33.5 50.1 3.3 5.0 95 65.459.6 6.5 6.0 10% Ethanol 25 19.3 12.6 1.9 1.3 95 80.9 70.8 8.1 7.1 40%Ethanol 25 23.9 27.9 2.4 2.8 95 91.6 64.6 9.2 6.5 pH 2 Buffer 25 56.132.7 5.6 3.3 95 67.5 73.2 6.8 7.3 pH 4 Buffer 25 8.8 8.1 0.9 0.8 95 27.824.2 2.8 2.4 pH 7 Buffer 25 10.8 14.7 1.1 1.5 95 41.3 40.0 4.1 4.0 pH 10Buffer 25 38.5 38.3 3.9 3.8 95 83.9 65.4 8.4 6.5

Although there was considerable variability among replicates, generally,the average amount of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid release (% recovery) was comparable between ground and intacttablets. As expected, extraction at near boiling conditions (95° C.)increased drug release. The most efficient solvents for extraction(>50%) of ground tablets at 25° C. were vinegar and pH 2 buffer. Atelevated temperature, the solvents that allowed >50% recovery for groundproduct were vinegar, 0.1M HCl, 10% ethanol, 40% ethanol, pH 2 buffer,and pH 10 buffer. Extract concentrations of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid from ground and intact tablets across all conditions ranged from0.8 mg/mL to 9.2 mg/mL.

F. Organic Solvent Extraction

This series of extractions was performed in a similar manner as theaqueous solvent extractions previously described. Ground and intacttablets were extracted in 10 mL of solvent for 10 minutes on an orbitalshaker at 100 rpm. Aliquots were removed and filtered with 0.45 μm PTFEfilters for HPLC analysis. The solvents utilized in these assessmentswere 95% ethanol, isopropyl alcohol, acetone and hexane.

Result Summary and Discussion

Organic solvent extraction of tablets was inefficient (<10%) across allconditions. The most efficient solvent utilized in this assessment was95% ethanol that provided an average % recovery of 8.2%5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid (range, 6.0%-10.3%).

G. Syringe Evaluation

Extracted solutions remaining from the aqueous extractions were assessedfor syringeability. In this assessment, a 10 mL syringe fitted with a 25gauge needed was utilized to withdraw as much of the remaining 7 mLvolume of extracted solution as possible into the syringe. A cotton ballwas used to filter the solution by inserting the needle in the cottonball during aspiration of the solution. The volume of solutionsuccessfully aspirated into the syringe was estimated and an aliquot wastested to determine the milligram amount of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenye-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid loaded into the syringe.

Result Summary and Discussion

Of the remaining 7 mL volume of solution remaining from extraction of apowdered tablet, the typical volume that could be loaded into a 10 mLsyringe fitted with a 25 gauge needle (cotton ball filtration) was 4-5mL of solution. Thus, the loss of available solution ranged fromapproximately 30% to 43% in this simulated assessment of preparing asolution for intravenous injection. The absolute amount of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid varied with the type of solvent. Overall, there was no evidence ofresistance to syringeability in these assessments.

H. Multiple Tablet Extraction

This initial series of extractions was designed to assess the extractionefficiency of selected aqueous-based solvents for preparation of tabletsfor injection. The extractions were performed with water and 0.1M HCl onground tablets with equivalent weight of 2 and 4 tablets. The extractionwas conducted with 10 mL of solvent for 10 minutes on an orbital shakerat 100 rpm at 95° C. Following extraction, aliquots were removed andfiltered with 0.45 μm PTFE filters for HPLC analysis. The rationale forthe design of this study was that individuals might attempt to extract alarger dose of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid through the use of heat, larger volumes and possibly use of acidicmedia. Although 10 mL volume is large for injection purposes, it isplausible that such a volume might be attempted, especially given thelimited solubility of the5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid. Also, it should be noted that although injection of a highly acidsolution (0.1M HCl) would likely result in discomfort and tissue injury,some individuals may attempt such procedures.

Result Summary and Discussion

The mean extraction efficiencies and concentration of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid in this extraction assessment are shown in Table 5.

TABLE 5 Mean extraction efficiencies of active ingredient in extractionassessment Number of Tablet Concentration, Solvent Equivalents %Recovery mg/mL Water 2 41.2 8.2 4 22.4 9.0 0.1M HCl 2 76.8 15.4 4 46.118.5

It would appear that both water and 0.1M HCl have nearly reachedcapacity for dissolution of the5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid under the two tablet equivalent extraction condition at 95° C. Theeffect of doubling the amount of potentially extractable5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid to four equivalents provided nearly equal concentrations of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid in both water and acid solution. At the same time, % recovery forthe 4 tablet condition was approximately one-half of the two tabletcondition indicating each solvent had reached a capacity limitingcondition.

I. Large Volume Extraction

Given that the5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid exhibits limited solubility in some solvents, extraction with alarger volume might be attempted by individuals attempting to isolatethe5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid for administration by non-intended routes. This extractionassessment was performed with ground and intact tablets. Extractionswere conducted with 30 mL volume of solvents (water, 0.1M HCl, 10%ethanol, and 95% ethanol) at 25° C. for 10 minutes and 24 hours on anorbital shaker at 100 rpm, then filtered with a 0.45 μm PTFE filter forHPLC analysis.

Result Summary and Discussion

The mean extraction efficiencies and concentration of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid in this extraction assessment are shown in Table 6 below.

TABLE 6 Extraction efficiencies of active ingredient in extractionassessment % % Conc. Conc. Extraction Recovery, Recovery, (mg/mL),(mg/mL), Solvent Time Ground Intact Ground Intact Water 10 min 19.8 0.90.7 0.03 24 hr 22.1 18.4 0.8 0.7 0.1M HCl 10 min 67.6 2.9 2.3 0.1 24 hr68.1 35.9 2.5 1.3 10% Ethanol 10 min 26.7 5.1 0.9 0.2 24 hr 28.8 47.71.1 1.8 95% Ethanol 10 min 27.0 0.06 0.9 0.0 24 hr 29.9 5.8 1.1 0.2

There was little difference in percent recovery of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid from the ground tablet following a 10 minute extraction period ascompared to extending the extraction period to 24 hours. In contrast,extraction of intact tablets for 10 minutes was considerably lessefficient than for 24 hours. It is likely that the increased surfacearea of the ground material was responsible for the rapidness of the 10minute extraction outcome. All concentrations of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]amino}-methyl)-2-methoxy-benzoicacid in the different solvent extracts were consistently <3 mg/mL.

J. Small Volume Extraction

Smaller volume extractions are commonly used by individuals attemptingto prepare a solution for injection. Water and saline are commonly usedsolvents for injection and were included in this study. Although itseems unlikely that 0.1M HCl would be used as a solvent because of ittoxic effects, it was included as a “worst-case scenario” inconsideration of the solubility properties of the5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid in the tablets. This extraction assessment was performed withground and intact tablets. Extractions were conducted with 5 mL volumeof solvents (water, saline, and 0.1M HCl) at 25° C. and 95° C. for 10minutes and 24 hours on an orbital shaker at 100 rpm, then filtered witha 0.45 μm PTFE filter for HPLC analysis.

Result Summary and Discussion

The mean extraction efficiencies and concentration of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid in this extraction assessment are shown in Table 7 below.

TABLE 7 Extraction efficiencies of active ingredient in extractionassessment Conc. Conc. Extraction % Recovery, % Recovery, (mg/mL),(mg/mL), Solvent Temp., ° C. Time Ground Intact Ground Intact Water 2510 min 16.2 16.4 3.2 3.3 95 10 min 34.7 32.4 6.9 6.5 25 24 hr 17.1 17.23.4 3.5 95 24 hr 28.3 25.9 5.7 5.2 Saline 25 10 min 16.9 17.5 3.4 3.5 9510 min 46.2 41.0 9.2 8.2 25 24 hr 17.6 17.7 3.5 3.6 95 24 hr 24.1 23.54.8 4.7 0.1M HCl 25 10 min 100.6 86.2 20.1 17.2 95 10 min 90.2 102.318.1 20.5 25 24 hr 91.8 95.1 18.4 19.0 95 24 hr 79.1 83.9 15.8 16.8

Increasing the extraction temperature from 25° C. to 95° C. enhancedrecovery and solubility of the5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid in water and saline; however extending extraction time did notappear to extract additional drug. Concentration of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid in water and saline was consistently <10 mg/mL. The use of 0.1M HClsubstantially enhanced recovery (nearly quantitative) and concentrationof5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenye-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid in solution although extraction for 24 hr at 95° C. appeared toproduce loss of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid (decomposition). Concentrations of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid in the acid extracts were in the range of 16-21 mg/mL.

K. Filter Evaluation

With the discovery that suspensions of material containing5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid were falsely elevating measures of drug release and recovery, anassessment was performed to determine how different types of filtrationsystems might affect drug release measurements. Extractions wereperformed on ground and intact tablets. Extractions were conducted with5 mL volume of solvent (water, 0.1M HCl, 10% ethanol and 95% ethanol) at25° C. for 10 minutes on an orbital shaker at 100 rpm. Followingextraction, extracts were filtered as follows: unfiltered (control);0.45 μm PTFE filter; cotton ball filter; cigarette filter; and coffeepaper filter.

Result Summary and Discussion

The mean extraction efficiencies and concentration of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid in this extraction assessment are shown in Table 8 below.

Table 8. Extraction efficiencies of active ingredient in extractionassessment

TABLE 8 Extraction efficiencies of active ingredient in extractionassessment % Re- % Re- Conc. Conc. covery, covery, (mg/mL), (mg/mL),Solvent Filter Type Ground Intact Ground Intact Water Unfiltered 49.161.9 9.8 12.4 0.45 μm PTFE filter 16.5 16.9 3.3 3.4 Cotton Ball 18.216.8 3.6 3.4 Cigarette Filter 18.5 16.9 3.7 3.4 Coffee Filter 16.8 17.13.4 3.4 0.1M Unfiltered 74.3 83.2 14.9 16.6 HCl 0.45 μm PTFE filter 76.787.4 15.3 17.5 Cotton Ball 56.6 86.3 11.3 17.3 Cigarette Filter 37.682.8 7.5 16.6 Coffee Filter 63.2 85.3 12.6 17.1 10% Unfiltered 90.7 76.418.1 15.3 Ethanol 0.45 μm PTFE filter 22.4 23.5 4.5 4.7 Cotton Ball 24.023.7 4.8 4.8 Cigarette Filter 24.9 24.4 5.0 4.9 22.6 22.9 4.5 4.6 95%Unfiltered 62.0 0.1 12.4 0.03 Ethanol 0.45 μm PTFE filter 7.3 0.1 1.50.02 Cotton Ball 26.7 0.04 5.3 0.01 Cigarette Filter 16.0 0.02 3.2 0.0Coffee Filter 10.4 0.03 2.1 0.01

Filtration had a substantial effect on apparent drug release inextractions with solvents that exhibit limited solubility (i.e., water,10% ethanol, 95% ethanol) for the5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid. Without filtration, the evidence appears persuasive that particlesuspension was the cause of these differences. When suspensions wereprepared for assay by HPLC, the5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid became soluble in the HPLC mobile phase and this accounted for theelevated “readings” (measurements). In contrast, extraction with 0.1MHCl showed little difference in drug release and recovery with orwithout filtration. Since the5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid exhibits considerably greater solubility in 0.1M HCl, there werenot suspended particles that would have been filtered out. It should benoted that this somewhat unusual behavior (forming particle suspensionsin solvents with limited solubility for the5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoic acid) adds a source of considerablevariability that would be highly dependent upon conditions employedduring extraction. This unique property of the formulation is notexpected to be easily discoverable by individuals that might attemptextraction with common household solvents.

The sustained amount of time that an extraction solvent is in contactwith a formulation can substantially influence how much active isreleased (and dissolved in the extraction solvent). This is particularlytrue for actives that exhibit limited solubility in the chosen solvent.Review of data generated in the study involving extraction with 30 mL ofsolvent for 10 minutes and 24 hours generally indicated greatervariability (% RSD) for the shorter extraction time compared to the 24hour extraction. To assess the influence to extraction time, a detailedtime course study was conducted to determine the impact of extractiontime on data variability and on percent recovery of the5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid from the tablets.

Extractions were performed on single ground and intact tablets (75 mgstrength). Extractions were conducted with 22.5 mL volume of solvent(water, 0.1M HCl, vinegar, and pH 7 buffer) at 25° C. and 95° C. on anorbital shaker at 100 rpm. A 2 mL aliquot was withdrawn at each timepoint. The time points were as follows: 10, 20, 30, 45 and 60 minutes,4, 12, and 24 hours. Extracts were filtered with a 0.45 μm PTFE filterfor HPLC analyses.

Result Summary and Discussion

The mean extraction efficiencies of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid over time and % RSD (n=⁻6 replicates) in this extraction assessmentare shown in the Table 9, below.

TABLE 9 Extraction efficiencies of active ingredient over time TimeVessel 10 20 30 45 1 4 12 24 minutes minutes minutes minutes Hour HoursHours Hours Amount of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoic acid releasedlabel claim (%) Water Ground—25° C. Mean 27.21 30.10 31.45 33.55 35.5144.16 53.07 59.37 RSD 28.6 21.7 19.0 21.7 16.3 15.7 13.5 11.0 Amount of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoic acid releasedlabel claim (%) Water Whole—25° C. Mean 2.04 12.59 18.75 20.86 25.4036.81 51.21 55.07 RSD 43.8 63.7 32.8 38.2 24.6 11.8 4.6 4.0 Amount of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoic acid releasedlabel claim (%) Water Ground—95° C. Mean 56.90 64.63 70.42 75.04 81.2791.07 94.43 94.42 RSD 45.1 33.9 26.6 20.6 14.4 4.4 3.6 3.0 Amount of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoic acid releasedlabel claim (%) Water Whole—95° C. Mean 80.78 83.66 84.16 87.39 89.4393.21 93.11 93.32 RSD 6.5 5.8 5.1 5.1 4.8 3.3 3.9 3.0 Amount of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoic acid releasedlabel claim (%) 0.1M HCl Ground—25° C. Mean 87.33 89.02 91.07 93.5092.99 97.84 101.50 101.39 RSD 30.5 30.2 24.7 17.2 18.3 7.3 1.9 2.0Amount of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoic acid releasedlabel claim (%) 0.1M HCl Whole—25° C. Mean 2.69 14.32 31.30 35.88 59.1677.52 101.48 102.69 RSD 51.7 72.2 44.5 21.8 19.5 9.4 1.9 2.8 Amount of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoic acid releasedlabel claim (%) 0.1M HCl Ground—95° C. Mean 99.59 95.96 97.03 96.9697.30 94.15 86.08 77.39 RSD 1.9 2.1 2.0 2.2 2.3 2.3 2.4 2.5 Amount of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoic acid releasedlabel claim (%) 0.1M HCl Whole—95° C. Mean 48.15 71.95 81.70 89.30 93.49100.44 93.17 82.24 RSD 17.7 8.3 5.1 4.6 2.9 3.1 1.8 2.6 Amount of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoic acid releasedlabel claim (%) Vinegar Ground—25° C. Mean 88.42 88.76 90.61 91.82 93.9398.38 100.89 101.12 RSD 3.6 4.2 3.3 4.3 5.3 2.5 1.8 1.4 Amount of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoic acid releasedlabel claim (%) Vinegar Whole—25° C. Mean 17.12 18.23 20.04 35.03 42.7258.04 101.52 105.46 RSD 207.3 171.5 158.6 97.4 67.5 36.9 5.3 2.7 Amountof5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoic acid releasedlabel claim (%) Vinegar Ground—95° C. Mean 94.28 98.54 100.73 101.69103.65 105.12 104.81 102.88 RSD 11.0 8.6 7.3 5.4 4.6 4.0 4.5 4.4 Amountof5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoic acid releasedlabel claim (%) Vinegar Whole—95° C. Mean 40.62 55.03 83.87 91.96 97.91111.12 111.29 109.85 RSD 4.1 8.9 23.7 16.5 10.8 4.7 4.5 5.6 Amount of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoic acid releasedlabel claim (%) pH 7 Ground—25° C. Mean 39.81 44.07 47.83 50.48 52.7659.47 68.50 76.49 RSD 7.5 10.7 16.1 15.7 15.1 14.4 8.8 7.9 Amount of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoic acid releasedlabel claim (%) pH 7 Whole—25° C. Mean 3.50 6.21 9.15 10.25 13.68 22.7442.44 54.14 RSD 32.5 67.3 65.5 41.9 15.7 18.7 16.3 10.0 Amount of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoic acid releasedlabel claim (%) pH 7 Ground—95° C. Mean 58.27 77.50 82.71 87.58 90.9998.74 98.87 96.37 RSD 23.3 13.8 9.6 5.7 4.3 5.4 4.9 5.0 Amount of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoic acid releasedlabel claim (%) pH 7 Whole—95° C. Mean 51.73 67.29 74.12 77.83 81.7097.02 100.80 97.68 RSD 17.1 4.3 7.4 2.4 2.8 4.0 3.4 3.5

An analysis of the mean % RSD from these data across the four solventsis shown in the Table 10 below.

TABLE 10 Mean % RSD across the four solvents Time, Mean % RSD Mean % RSDMean % RSD Mean % RSD hr 25° C. Ground 25° C. Whole 95° C. Ground 95° C.Whole 0.17 17.6 83.8 20.3 11.4 0.33 16.7 93.7 14.6 6.8 0.5 15.8 75.411.4 10.3 0.75 14.7 49.8 8.5 7.2 1 13.8 31.8 6.4 5.3 4 10.0 19.2 4.0 3.812 6.5 7.0 3.9 3.4 24 5.6 4.9 3.7 3.7

In general, there was greater variability in extraction of the5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid from the tablets with shorter times and at the lower extractiontemperature (25° C.) compared to 95° C. Also, there was less variabilityin extraction of the ground tablet at 25° C. compared to the wholetablet.

Representative time course plots of cumulative percent recovery of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenye-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid under different extraction conditions are shown in FIG. 2.

The abbreviations used in the plots are as follows:

Gr=ground tablet; Wh =whole, intact tablet.General observations from these time course data include the following:

-   -   With short extraction times (e.g., 10 minutes), ground tablets        are more efficiently extracted than whole tablets    -   With extended extraction time (e.g., 24 hours), there is little        difference in extractability of        5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoic        acid from ground versus whole tablets    -   Extraction under heated conditions (e.g., 95° C.) substantially        increases % recovery at shorter extraction times    -   Acidic solvents are more efficient in extracting        5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenye-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoic        acid than water or pH neutral solvents

L. Investigations of Multiple Tablet Extractions

In addition to extractions of multiple tablets, additional experimentswere conducted to determine the optimal conditions for preparation ofmultiple tablets for injection. Initially, attempts were made toidentify the correct combination(s) of solvent and ground tablets thatwould yield the most concentrated recoverable solution of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid from tablets. Based on these initial assessments, a study wasdesigned in which extractions were conducted with powdered tablets at25° C. and 95° C. with the equivalent of four tablets. The extractionswere conducted for 10 minutes, one hour, and 12 hours with 10 mL ofwater, vinegar, 0.1M HCl, and saline. The extraction vials were shakenon an orbital shaker at 200 rpm. Following extraction, extracts werefiltered with a 0.45 μm PTFE filter for HPLC analyses.

Result Summary and Discussion

The mean extraction efficiencies and concentrations of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid over time in this extraction assessment are shown in Table 11below. Data for the four solvents are listed in order from lowest(water) to highest polarity and acidity (0.1M HCl).

TABLE 11 Extraction efficiencies and concentrations of 5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2- methoxy-benzoicacid over time % Conc. % Conc. Extraction Recovery, (mg/mL), Recovery,(mg/mL), Solvent Time 25° C. 25° C. 95° C. 95° C. Water 10 min 8.1 3.324.2 9.7  1 hr 9.5 3.8 23.6 9.4 12 hr 8.9 3.6 25.1 10.0 Saline 10 min8.9 3.5 18.5 7.4  1 hr 9.8 3.9 22.3 8.9 12 hr 9.8 3.9 24.1 9.6 Vinegar10 min 5.6 2.2 16.0 6.4  1 hr 27.7 11.1 48.0 19.2 12 hr 78.8 31.5 79.531.8 0.1M HCl 10 min 18.0 7.2 34.3 13.7  1 hr 39.7 15.9 70.2 28.1 12 hr88.3 35.3 90.6 36.3

These data suggest that the maximal concentration of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid that can be extracted with water and saline is approximately 4mg/mL at room temperature (e.g., 25° C.) and approximately 10 mg/mLunder near boiling conditions (e.g., 95° C.). Further, there appeared tobe little difference in maximal concentrations with water and salineregardless of extraction time. Use of an acidic solvent such as vinegarand the highly acidic solvent, 0.1MHCl led to the production of moreconcentrated extracts of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenye-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid with maximal values of approximately 36 mg/mL being achieved with a12 hour extraction period. There was both time and temperaturedependence on concentration with these two solvents. With vinegar,maximal concentrations over the 10 minute and one hour extraction periodwere approximately 2-11 mg/mL at 25° C. and 6-19 mg/mL, respectively.With 0.1M HCl, maximal concentrations over the 10 minute and one hourextraction period were approximately 7-16 mg/mL at 25° C. and 14-28mg/mL, respectively. Given these maximal concentrations, an estimate ofthe required volume of extracted5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid that would deliver 350 mg of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid by injection is shown in Table 12 below.

TABLE 12 Estimate of the required volume of extracted5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoic acid that would deliver 350 mgof active ingredient by injection. Injection Volume Injection Volume(mL) Required for (mL) Required for 350 mg of 5-({[2- 350 mg of 5-({[2-Amino-3-(4- Amino-3-(4- carbamoyl-2,6- carbamoyl-2,6- dimethyl-phenyl)-dimethyl-phenyl)- propionyl]-[1-(4- propionyl]-[1-(4- phenyl-1H-phenyl-1H- imidazol-2-yl)- imidazol-2-yl)- ethyl]-amino}- ethyl]-amino}-Conc. methyl)-2- Conc. methyl)-2- Extraction (mg/mL), methoxy-benzoic(mg/mL), methoxy-benzoic Solvent Time 25° C. acid at 25° C. 95° C. acidat 95° C. Water 10 min 3.3 106.1 9.7 36.1  1 hr 3.8 92.1 9.4 37.2 12 hr3.6 97.2 10 35.0 Saline 10 min 3.5 100.0 7.4 47.3  1 hr 3.9 89.7 8.939.3 12 hr 3.9 89.7 9.6 36.5 Vinegar 10 min 2.2 159.1 6.4 54.7  1 hr11.1 31.5 19.2 18.2 12 hr 31.5 11.1 31.8 11.0 0.1M HCl 10 min 7.2 48.613.7 25.5  1 hr 15.9 22.0 28.1 12.5 12 hr 35.3 9.9 36.3 9.6

Typical opioid injection volumes are generally in the 1-3 mL range butplausibly could be as high as 10 mL. Assuming that a 350 mg injecteddose of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid would be required to produce a euphoric effect, it does not appearfeasible that tablets could be prepared for injection with sufficientcontent of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid when using typical injection solvents (water, saline). Although asufficiently concentrated solution of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid by extraction does appear to be possible through the use of vinegarand 0.1M HCl, it is unlikely that many individuals would be willing toaccept the potential toxic risks that these solvents present.

The foregoing examples of the present disclosure have been presented forpurposes of illustration and description. Furthermore, these examplesare not intended to limit the disclosure to the form disclosed herein.Consequently, variations and modifications commensurate with theteachings of the description of the disclosure, and the skill orknowledge of the relevant art, are within the scope of the presentdisclosure. The specific embodiments described in the examples providedherein are intended to further explain the best mode known forpracticing the disclosure and to enable others skilled in the art toutilize the disclosure in such, or other, embodiments and with variousmodifications required by the particular applications or uses of thepresent disclosure. It is intended that the appended claims be construedto include alternative embodiments to the extent permitted by the priorart.

1. A solid pharmaceutical dosage formulation comprising about 20 mg/doseto about 200 mg/dose of5({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid, about 60-80% by weight of silicified microcrystalline cellulose,about 0.55-0.95% by weight of colloidal silicon dioxide, about 3-7% byweight of crospovidone, about 5-15% mannitol, and about 0.55-0.95% byweight of magnesium stearate, wherein extraction of the formulation ofwater or saline at 25° C. for up to 12 hours produces a concentration of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid of less than or approximately 4 mg/ml.
 2. The solid pharmaceuticaldosage formulation of claim 1, formulated as a tablet or as a capsule.3. The solid pharmaceutical dosage formulation of claim 1, furthercomprising a coating selected from a sugar coating, a gelatin coating, afilm coating, and an enteric coating.
 4. The solid pharmaceutical dosageformulation of claim 3, comprising about 75 mg to about 200 mg of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid.
 5. The solid pharmaceutical dosage formulation of claim 3,comprising about 75 mg of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid.
 6. The solid pharmaceutical dosage formulation of claim 3,comprising about 100 mg of 5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid.
 7. The solid pharmaceutical dosage formulation of claim 1, whereinthe formulation is substantially or completely free of naloxone. 8-29.(canceled)
 30. The solid pharmaceutical dosage formulation of claim 1,wherein the formulation comprises: about 65-75% by weight of silicifiedmicrocrystalline cellulose, about 0.65-0.85% by weight of colloidalsilica, about 7.5-12.5% by weight of mannitol, from about 4-6% by weightof crospovidone, and from about 0.65-0.85% by weight of magnesiumstearate.
 31. The solid pharmaceutical dosage formulation of claim 1,wherein the formulation comprises about 71% by weight of silicifiedmicrocrystalline cellulose, about 0.75% by weight of colloidal silica,about 10% by weight of mannitol, about 5% by weight of crospovidone, andabout 0.75% by weight of magnesium stearate.
 32. The solidpharmaceutical dosage formulation of claim 1, comprising a film coatingpresent at about 2-4% by weight of the pharmaceutical composition. 33.The pharmaceutical composition of claim 32, wherein the film coating ispresent at about 3% by weight of the pharmaceutical composition.
 34. Thepharmaceutical composition of claim 32 , wherein the film coating is apolyvinyl alcohol (PVA) based film coating.
 35. The pharmaceuticalcomposition of claim 1, further comprising at least one inertpharmaceutical excipient selected from a pharmaceutically acceptabledissolution-rate-modifying agent, a pharmaceutically acceptableplasticizer, a pharmaceutically acceptable coloring agent, apharmaceutically acceptable opacifier, pharmaceutically acceptableanti-oxidant, a pharmaceutically acceptable preservative, a flavorant, aneutralizing agent, a buffering agent and combinations thereof.
 36. Apharmaceutical composition comprising: about 75 mg of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid; about 426 mg of silicified microcrystalline cellulose; about 4.5mg of colloidal silica; about 60 mg of mannitol; about 30 mg ofcrospovidone; and about 4.5 mg by weight of magnesium stearate; whereinextraction of the composition with water or saline at 25° C. for up to12 hours produces a concentration of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid of less than or approximately 4 mg/ml.
 37. A pharmaceuticalcomposition comprising: about 100 mg of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid; about 568 mg of silicified microcrystalline cellulose; about 6 mgof colloidal silica; about 80 mg of mannitol; about 40 mg ofcrospovidone; and about 6 mg by weight of magnesium stearate; whereinextraction of the composition with water or saline at 25° C. for up to12 hours produces a concentration of5-({[2-Amino-3-(4-carbamoyl-2,6-dimethyl-phenyl)-propionyl]-[1-(4-phenyl-1H-imidazol-2-yl)-ethyl]-amino}-methyl)-2-methoxy-benzoicacid of less than or approximately 4 mg/ml.